A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain

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A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain. / Hughes, Eleri; Whittaker, Christopher A P; Barsukov, Igor L et al.
In: Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1808, No. 4, 04.2011, p. 1021-31.

Research output: Contribution to Journal/Magazine › Journal article › peer-review

Harvard

Hughes, E, Whittaker, CAP, Barsukov, IL, Esmann, M & Middleton, DA 2011, 'A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain', Biochimica et Biophysica Acta (BBA) - Biomembranes, vol. 1808, no. 4, pp. 1021-31. https://doi.org/10.1016/j.bbamem.2010.11.024

APA

Hughes, E., Whittaker, C. A. P., Barsukov, I. L., Esmann, M., & Middleton, D. A. (2011). A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1808(4), 1021-31. https://doi.org/10.1016/j.bbamem.2010.11.024

Vancouver

Hughes E, Whittaker CAP, Barsukov IL, Esmann M, Middleton DA. A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain. Biochimica et Biophysica Acta (BBA) - Biomembranes. 2011 Apr;1808(4):1021-31. doi: 10.1016/j.bbamem.2010.11.024

Author

Hughes, Eleri ; Whittaker, Christopher A P ; Barsukov, Igor L et al. / A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain. In: Biochimica et Biophysica Acta (BBA) - Biomembranes. 2011 ; Vol. 1808, No. 4. pp. 1021-31.

Bibtex

@article{c36cffaa9ab3446e97706e7699f8fa02,

title = "A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain",

abstract = "Phospholemman (PLM) is a single-span transmembrane protein belonging to the FXYD family of proteins. PLM (or FXYD1) regulates the Na,K-ATPase (NKA) ion pump by altering its affinity for K(+) and Na(+) and by reducing its hydrolytic activity. Structural studies of PLM in anionic detergent micelles have suggested that the cytoplasmic domain, which alone can regulate NKA, forms a partial helix which is stabilized by interactions with the charged membrane surface. This work examines the membrane affinity and regulatory function of a 35-amino acid peptide (PLM(38-72)) representing the PLM cytoplasmic domain. Isothermal titration calorimetry and solid-state NMR measurements confirm that PLM(38-72) associates strongly with highly anionic phospholipid membranes, but the association is weakened substantially when the negative surface charge is reduced to a more physiologically relevant environment. Membrane interactions are also weakened when the peptide is phosphorylated at S68, one of the substrate sites for protein kinases. PLM(38-72) also lowers the maximal velocity of ATP hydrolysis (V(max)) by NKA, and phosphorylation of the peptide at S68 gives rise to a partial recovery of V(max). These results suggest that the PLM cytoplasmic domain populates NKA-associated and membrane-associated states in dynamic equilibrium and that phosphorylation may alter the position of the equilibrium. Interestingly, peptides representing the cytoplasmic domains of two other FXYD proteins, Mat-8 (FXYD3) and CHIF (FXYD4), have little or no interaction with highly anionic phospholipid membranes and have no effect on NKA function. This suggests that the functional and physical properties of PLM are not conserved across the entire FXYD family.",

keywords = "Adenosine Triphosphate, Amino Acid Sequence, Animals, Binding, Competitive, Calorimetry, Cell Membrane, Circular Dichroism, Humans, Kinetics, Lipid Bilayers, Magnetic Resonance Spectroscopy, Membrane Proteins, Molecular Sequence Data, Neoplasm Proteins, Peptide Fragments, Phospholipids, Phosphoproteins, Phosphorylation, Potassium, Protein Binding, Serine, Sodium, Sodium-Potassium-Exchanging ATPase",

author = "Eleri Hughes and Whittaker, {Christopher A P} and Barsukov, {Igor L} and Mikael Esmann and Middleton, {David A}",

note = "Copyright {\textcopyright} 2010. Published by Elsevier B.V.",

year = "2011",

month = apr,

doi = "10.1016/j.bbamem.2010.11.024",

language = "English",

volume = "1808",

pages = "1021--31",

journal = "Biochimica et Biophysica Acta (BBA) - Biomembranes",

issn = "0005-2736",

publisher = "Elsevier",

number = "4",

}

RIS

TY - JOUR

T1 - A study of the membrane association and regulatory effect of the phospholemman cytoplasmic domain

AU - Hughes, Eleri

AU - Whittaker, Christopher A P

AU - Barsukov, Igor L

AU - Esmann, Mikael

AU - Middleton, David A

PY - 2011/4

Y1 - 2011/4

N2 - Phospholemman (PLM) is a single-span transmembrane protein belonging to the FXYD family of proteins. PLM (or FXYD1) regulates the Na,K-ATPase (NKA) ion pump by altering its affinity for K(+) and Na(+) and by reducing its hydrolytic activity. Structural studies of PLM in anionic detergent micelles have suggested that the cytoplasmic domain, which alone can regulate NKA, forms a partial helix which is stabilized by interactions with the charged membrane surface. This work examines the membrane affinity and regulatory function of a 35-amino acid peptide (PLM(38-72)) representing the PLM cytoplasmic domain. Isothermal titration calorimetry and solid-state NMR measurements confirm that PLM(38-72) associates strongly with highly anionic phospholipid membranes, but the association is weakened substantially when the negative surface charge is reduced to a more physiologically relevant environment. Membrane interactions are also weakened when the peptide is phosphorylated at S68, one of the substrate sites for protein kinases. PLM(38-72) also lowers the maximal velocity of ATP hydrolysis (V(max)) by NKA, and phosphorylation of the peptide at S68 gives rise to a partial recovery of V(max). These results suggest that the PLM cytoplasmic domain populates NKA-associated and membrane-associated states in dynamic equilibrium and that phosphorylation may alter the position of the equilibrium. Interestingly, peptides representing the cytoplasmic domains of two other FXYD proteins, Mat-8 (FXYD3) and CHIF (FXYD4), have little or no interaction with highly anionic phospholipid membranes and have no effect on NKA function. This suggests that the functional and physical properties of PLM are not conserved across the entire FXYD family.

AB - Phospholemman (PLM) is a single-span transmembrane protein belonging to the FXYD family of proteins. PLM (or FXYD1) regulates the Na,K-ATPase (NKA) ion pump by altering its affinity for K(+) and Na(+) and by reducing its hydrolytic activity. Structural studies of PLM in anionic detergent micelles have suggested that the cytoplasmic domain, which alone can regulate NKA, forms a partial helix which is stabilized by interactions with the charged membrane surface. This work examines the membrane affinity and regulatory function of a 35-amino acid peptide (PLM(38-72)) representing the PLM cytoplasmic domain. Isothermal titration calorimetry and solid-state NMR measurements confirm that PLM(38-72) associates strongly with highly anionic phospholipid membranes, but the association is weakened substantially when the negative surface charge is reduced to a more physiologically relevant environment. Membrane interactions are also weakened when the peptide is phosphorylated at S68, one of the substrate sites for protein kinases. PLM(38-72) also lowers the maximal velocity of ATP hydrolysis (V(max)) by NKA, and phosphorylation of the peptide at S68 gives rise to a partial recovery of V(max). These results suggest that the PLM cytoplasmic domain populates NKA-associated and membrane-associated states in dynamic equilibrium and that phosphorylation may alter the position of the equilibrium. Interestingly, peptides representing the cytoplasmic domains of two other FXYD proteins, Mat-8 (FXYD3) and CHIF (FXYD4), have little or no interaction with highly anionic phospholipid membranes and have no effect on NKA function. This suggests that the functional and physical properties of PLM are not conserved across the entire FXYD family.

KW - Adenosine Triphosphate

KW - Amino Acid Sequence

KW - Animals

KW - Binding, Competitive

KW - Calorimetry

KW - Cell Membrane

KW - Circular Dichroism

KW - Humans

KW - Kinetics

KW - Lipid Bilayers

KW - Magnetic Resonance Spectroscopy

KW - Membrane Proteins

KW - Molecular Sequence Data

KW - Neoplasm Proteins

KW - Peptide Fragments

KW - Phospholipids

KW - Phosphoproteins

KW - Phosphorylation

KW - Potassium

KW - Protein Binding

KW - Serine

KW - Sodium

KW - Sodium-Potassium-Exchanging ATPase

U2 - 10.1016/j.bbamem.2010.11.024

DO - 10.1016/j.bbamem.2010.11.024

M3 - Journal article

C2 - 21130070

VL - 1808

SP - 1021

EP - 1031

JO - Biochimica et Biophysica Acta (BBA) - Biomembranes

JF - Biochimica et Biophysica Acta (BBA) - Biomembranes

SN - 0005-2736

IS - 4

ER -

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